The action of dexamethasone is initiated by, and strictly dependent upon,

The action of dexamethasone is initiated by, and strictly dependent upon, the interaction of the drug with its receptor followed by its translocation into the nucleus where modulates gene expression. the transcription process [22,23], and act as platform for Vitamin D3CVitamin D3 receptor interaction, inducing embryonic hippocampal cell differentiation [24]. We aimed to study the localization of the Dex in NLM after its translocation Rosuvastatin inside the nucleus and its effect in human lymphoblastic lymphoma T cell growth. 2. Results 2.1. Non-Hodgkins T Cell Human Lymphoblastic Lymphoma Cell Growth Is Suppressed by Dexamethasone We first investigated the effect of Dex on the non-Hodgkins T cell human lymphoblastic lymphoma cell line (SUP-T1). The results showed that in the control cells the specific activity of the DNA, calculated as cpm/g DNA, increased at 12 h, and reached a peak at 24 h, which corresponded to the S phase of the Rosuvastatin cell cycle (Figure 1a). Dex treatment caused a strong decrease of 3H-thymidine already detectable at 12 h; the value remained constant until 48 h, when it slightly increased (Figure 1a). At this time, the number of control cells was 278 13 number/mL and that of Dex-treated cells was 168 16. The difference in cell growth between control and Rosuvastatin experimental samples increased in time (Figure 1b). To highlight the inhibition of the cell cycle, we study the gene expression of [10,11,12]. Glyceraldehyde 3-phosphate dehydrogenase Rosuvastatin (GAPDH) has long been used as a default reference gene in quantitative mRNA profiling experiments. It is a ubiquitous enzyme that catalyzes the sixth step of glycolysis and plays a role in the control of gene expression and redox post-translational modifications [25]. Thus, its expression varied in response to a range of pathophysiological variables [25,26]. Our results showed that Dex treatment increased 8 times GAPDH expression (Figure 2). On the other hand, treatment with drugs changes the expression of many genes and a specific study is needed to identify the housekeeping genes [27]. Since at this moment no specific studies have been done on MPO SUP-T1 cells treated with Dex to identify housekeeping gene, we evaluated mRNA expression of Dex-treated cells in relation to mRNA of control cells, according to Schmittgen and Livak [28]. Our results showed that the massive block of proliferation was accompanied, at 24 h from Dex incubation, by an up-regulation of CDKN1A, CDKN1B and GADD45A equal to 4.27, 5.71 and 4.91 times respectively, in comparison with control samples (Figure 2). Since it has been demonstrated that Dex inhibited phospho signal transducer and activator of transcription 3 (phosphoSTAT3) [29], we performed experiments of immunoblotting after 24 h of drug incubation showing a strong decrease of STAT3 and phosphoSTAT3 content (Figure 3a,b). As control for immunoblotting technique we used -actin, normally used, but it increased strongly (Figure 3a). On the other hand it is known that Dex acted on actin networks [30]. Therefore it was not a good control but its increase was an indication that the reduction of STAT3 and phosphoSTAT3 was not due to the experimental defect. We next wondered whether later, at 48 h after Dex treatment, there might be changes in cell morphology and anti-apoptotic Bcl-2 protein content, up-regulated in T-cell acute lymphoblastic leukemia [31]. Hematoxylin-eosin staining showed round cells with nuclei intensely colored and cells ready for division. In the experimental sample it was possible to notice a change in the shape of the cells (Figure 4a). As shown in Figure 4b, Dex-treated cells were reduced in number but were bigger than controls with a small amount of the cells with altered morphology, similar to that indicated in Figure 4a. The percentage of Bcl-2 positive Rosuvastatin cells was reduced 5 times; the only positive cells were those that were ready for starting mitosis (Figure 4b,c). Figure 1 Effect of Dexamethasone on DNA synthesis and cell growth. (a) Incorporation of 3H-thymidine in non-Hodgkins T cell human lymphoblastic lymphoma cell line (SUP-T1) cells. Cells were cultured for 48 h without (C = control sample) or with 100 Dex. … Figure 2 Effect of Dexamethasone on.